Method for constructing mouse model for conditional knockout of CCR5 gene of endothelial cell

A technology of endothelial cells and construction methods, applied in the field of construction of endothelial cell conditional knockout of CCR5 gene mouse model, can solve the problem of no one carrying out specific knockout of CCR5

Active Publication Date: 2018-03-30
SHANDONG KEYUAN PHARMA
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  • Claims
  • Application Information

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Problems solved by technology

[0003] Based on the specific and unique regulatory mechanism and specific functional activity of CCR5 in different cells, it is necess

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  • Method for constructing mouse model for conditional knockout of CCR5 gene of endothelial cell
  • Method for constructing mouse model for conditional knockout of CCR5 gene of endothelial cell
  • Method for constructing mouse model for conditional knockout of CCR5 gene of endothelial cell

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Embodiment Construction

[0026] The present invention will be further described below in conjunction with embodiment. However, the scope of the present invention is not limited to the following examples. Those skilled in the art can understand that various changes and modifications can be made in the present invention without departing from the spirit and scope of the present invention.

[0027]The present invention provides general and / or specific descriptions of the materials and test methods used in the tests. While many of the materials and methods of manipulation which are employed for the purposes of the invention are well known in the art, the invention has been described here in as much detail as possible.

[0028] For all of the following examples, standard manipulations and purification methods known to those skilled in the art can be used. All temperatures are in °C (degrees Celsius) unless otherwise indicated. All reactions were performed at room temperature unless otherwise stated. Th...

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Abstract

The invention discloses a method for constructing a mouse model for conditional knockout of CCR5 gene of endothelial cell. The method comprises the following steps of firstly, obtaining a CCR5loxp/loxp mouse, and mating with a Tie-2-cre/ERT2 mouse, so as to obtain heterozygous mice with specific knockout of CCR5 gene of the endothelial cell; mutually mating the heterozygous mice, so as to obtain homozygous mice with specific knockout of CCR5 gene out of the endothelial cell. The constructing method disclosed by the invention has the advantages that the mouse with conditional knockout of endothelial cell gene is constructed via the CRISPR/Cas9 system, the mutation is led into the mouse by a cell specificity method, the missing of the CCR5 target gene occurs at the certain tissue organ of the test animal, the controllability of mechanism study is furthest realized, and the disadvantages that different tissues or cells are not distinguished, and target genes in all tissues or cells of themouse body are all removed in the conventional gene knockout technique is overcome.

Description

technical field [0001] The invention relates to a method for constructing an endothelial cell conditional knockout CCR5 gene mouse model, which belongs to the field of animal models and applications thereof. Background technique [0002] Chemokine receptor CCR5 (chemokine receptor 5) is a kind of G protein 7 transmembrane receptors, with a molecular weight of about 40.6KDa, composed of 352 amino acid residues, structurally can be divided into: extracellular N-terminal, 3 Extracellular loop regions (ECL1, ECL2, ECL3), 3 intracellular loop regions, 7 transmembrane α-helices and intracellular C-terminal (Szpakowska M, Perez Bercoff D, Chevigne A. Closing the ring: a fourthextracellular loop in chemokine receptors.Science signaling, 2014,7(341):pe21.), mainly expressed in monocytes and certain T cell subsets. The ligands of CCR5 are mainly chemokine CCL3 (chemokine C-C motif ligand 3), CCL4 (chemokine C-C motif ligand 4) and CCL5 (chemokine C-C motif ligand 5). As a G protein-...

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Application Information

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IPC IPC(8): C12N15/85C12N15/90A01K67/027
CPCA01K67/0276A01K2217/075A01K2227/105A01K2267/0362C07K14/47C12N15/8509C12N15/907
Inventor 张钟文廖琳董建军吴红霞张瀚允
Owner SHANDONG KEYUAN PHARMA
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